Bio-Rad Profinity Epoxide Resin User Manual

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Profinity

™

Epoxide Resin

Instruction Manual

Please read these instructions prior to using Profinity epoxide resin. If
you have any questions or comments regarding these instructions,
please contact your Bio-Rad Laboratories representative.

Table of Contents

Section 1 Product Description ................................................1

Section 2 General Considerations for

Ligand Coupling.......................................................3

Section 3 Column Packing.......................................................5

Section 4 Protein Binding and Elution ..................................7

Section 5 Renaturation of Eluted Proteins...........................8

Section 6 Assessing Protein Purity........................................8

Section 7 Regeneration and Storage.....................................9

Section 8 References................................................................9

Section 9 Ordering Information ..............................................9

Section 1

UNOsphere

O

OH

NHR (-SR, -OR)

R-NH2, -SH, -OH

UNOsphere

Product Description

Profinity epoxide is an activated macroporous resin for the immobilization of
various ligands of interest. Ligands containing amino, thiol, or hydroxyl groups can
be coupled to Profinity epoxide through an epoxy ring-opening reaction under mild
conditions.

Profinity epoxide is based on Bio-Rad’s proprietary and innovative UNOsphere

™

technology (US patent 6,423,666). Resins made using this technology have
properties of superb mechanical strength, open pore structure, optimized ligand
density, and low, nonspecific binding effects. These unique features of the
UNOsphere base matrix enable ligand-coupled Profinity resin to exhibit excellent
flow properties and to perform separations at very fast flow rates without
compromising protein binding, recovery, or purity. Profinity resin’s open-pore
structure is particularly useful for the purification of large biomolecules.

The base matrix of Profinity epoxide resin is stable across the entire pH range
(1–14) and is compatible with most reagents commonly used in protein
purification, such as denaturing agents, detergents, and reducing agents. It is
amenable to separations under native or denaturing conditions using liquid
chromatographic instrumentation, gravity flow columns, or sample preparation spin
columns.

The resin is supplied dry and is available in 5 g and 25 g quantities.

Note: UNOsphere support, from which Profinity epoxide is derived, was designed
to achieve the highest productivity possible (as measured in grams of target
molecule per operational hour per liter of support). UNOsphere media may be run
at the highest rates and loading capacities while staying within the pressure limits
of the column and chromatography system.

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Table 1: Characteristics of Profinity epoxide resin

Profinity epoxide

Functional group Epoxy group

Base matrix UNOsphere base matrix

Form Dry powder

Particle size 45–90 µm

Mean particle size 60 µm

Functional group density 50–132 µmol/g UNOsphere epoxide resin

Swelling factor (ml drained resin/g resin) 5.5–8.0

Recommended linear flow rate <600 cm/hr at 25°C

Maximum operating pressure (net)*

pH stability (base matrix of coupled resin)** 1–14
Chemical compatibility (base matrix

of coupled resin)**
Storage 4°C ambient temperature

Shelf life > 1 year at ambient temperature

Operational temperature 4–40°C
Autoclavability (base matrix
of coupled resin)**

**

Maximum pressure test: Profinity epoxide resin packed in a 1.1 x 30 cm Amicon column to a bed height
of 20 cm with 20 mM sodium phosphate buffer up to 43 psi (3 bar). Flow rates were increased stepwise
to 200 cm/h and held for 2 min at each step. The pressure-low curve for Profinity epoxide becomes
nonlinear at pressures above 80 psi.

****

Refers to base matrix of coupled resin. The stability of coupled ligands may be a limitation of an affinity

resin’s stability.

≥ 80 psi

Compatible with common buffers
and aqueous solutions

0.1 M sodium acetate at 120°C for 30 min

2